WEKO3
アイテム
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To detect abnormal epigenetic alterations in DNA methylation, convenient and cost-effective\nmethods are required for such research, in which multiple samples are processed simultaneously. We here present\nmethylated site display (MSD), a unique technique for the preparation of DNA libraries. By combining it with amplified\nfragment length polymorphism (AFLP) analysis, we developed a new method, MSD-AFLP.\nResults: Methylated site display libraries consist of only DNAs derived from DNA fragments that are CpG methylated\nat the 5′ end in the original genomic DNA sample. To test the effectiveness of this method, CpG methylation levels in\nliver, kidney, and hippocampal tissues of mice were compared to examine if MSD-AFLP can detect subtle differences\nin the levels of tissue-specific differentially methylated CpGs. As a result, many CpG sites suspected to be tissue-specific\ndifferentially methylated were detected. 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Methylated site display (MSD)-AFLP, a sensitive and affordable method for analysis of CpG methylation profiles
https://repo.qst.go.jp/records/47729
https://repo.qst.go.jp/records/47729c6729ca3-8812-4eb6-bf7a-d5e2fb53b97c
Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2017-04-20 | |||||
タイトル | ||||||
タイトル | Methylated site display (MSD)-AFLP, a sensitive and affordable method for analysis of CpG methylation profiles | |||||
言語 | ||||||
言語 | eng | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
資源タイプ | journal article | |||||
アクセス権 | ||||||
アクセス権 | metadata only access | |||||
アクセス権URI | http://purl.org/coar/access_right/c_14cb | |||||
著者 |
Aiba, Toshiki
× Aiba, Toshiki× Saito, Toshiyuki× Hayashi, Akiko× Sato, Shinji× Yunokawa, Harunobu× Maruyama, Toru× Fujibuchi, Wataru× Kurita, Hisaka× Tohyama, Chiharu× Ohsako, Seiichiroh× 相場 俊樹× 齋藤 俊行× 林 昭子 |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | Background: It has been pointed out that environmental factors or chemicals can cause diseases that are developmental in origin. To detect abnormal epigenetic alterations in DNA methylation, convenient and cost-effective methods are required for such research, in which multiple samples are processed simultaneously. We here present methylated site display (MSD), a unique technique for the preparation of DNA libraries. By combining it with amplified fragment length polymorphism (AFLP) analysis, we developed a new method, MSD-AFLP. Results: Methylated site display libraries consist of only DNAs derived from DNA fragments that are CpG methylated at the 5′ end in the original genomic DNA sample. To test the effectiveness of this method, CpG methylation levels in liver, kidney, and hippocampal tissues of mice were compared to examine if MSD-AFLP can detect subtle differences in the levels of tissue-specific differentially methylated CpGs. As a result, many CpG sites suspected to be tissue-specific differentially methylated were detected. Nucleotide sequences adjacent to these methyl-CpG sites were identified and we determined the methylation level by methylation-sensitive restriction endonuclease (MSRE)-PCR analysis to confirm the accuracy of AFLP analysis. The differences of the methylation level among tissues were almost identical among these methods. By MSD-AFLP analysis, we detected many CpGs showing less than 5% statistically significant tissue-specific difference and less than 10% degree of variability. Additionally, MSD-AFLP analysis could be used to identify CpG methylation sites in other organisms including humans. Conclusion: MSD-AFLP analysis can potentially be used to measure slight changes in CpG methylation level. Regarding the remarkable precision, sensitivity, and throughput of MSD-AFLP analysis studies, this method will be advantageous in a variety of epigenetics-based research. Keywords: DNA methylation profiling, AFLP, Epigenetics |
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書誌情報 |
BMC Molecular Biology 巻 18, 号 7, p. 1-11, 発行日 2017-03 |
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出版者 | ||||||
出版者 | BioMed Central | |||||
ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 1471-2199 | |||||
DOI | ||||||
識別子タイプ | DOI | |||||
関連識別子 | 10.1186/s12867-017-0083-2 | |||||
関連サイト | ||||||
識別子タイプ | URI | |||||
関連識別子 | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5345256/pdf/12867_2017_Article_83.pdf | |||||
関連名称 | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5345256/pdf/12867_2017_Article_83.pdf |